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標題: 莫三比克吳郭魚(Oreochromis mossambicus)適應在淡水、海水、去離子水環境中鰓上氯離子傳輸蛋白之分布
Differential distribution of chloride transporters in gills of fresh water-, seawater- and deionized water-acclimated euryhaline teleosts, Oreochromis mossambicus
作者: 趙廷偉
Chao, Ting-Wei
關鍵字: Oreochromis mossambicus;莫三比克吳郭魚;acclimate;deionized water;gill;chloride transporters;適應;去離子水;鰓;氯離子傳輸蛋白
出版社: 生命科學系所
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莫三比克吳郭魚(Oreochromis mossambicus)為廣鹽性硬骨魚類,可適應於淡水或海水。本實驗利用whole mount與雙重免疫螢光染色配合倒立式共軛焦雷射顯微鏡定性觀察,以瞭解莫三比克吳郭魚適應在不同離子濃度的環境下,其氯離子傳輸蛋白在鰓表皮細胞上的分布,藉此了解吳郭魚在適應不同環境時鰓表皮對氯離子的運輸機制。雙重免疫染色係以離子傳輸的主動力Na+ / K+ -ATPase(NKA)作對比染色,探討四種氯離子傳輸蛋白,包括Na+ / K+ / 2Cl- cotransporter(NKCC);Cystic fibrosis transmembrane regulator(CFTR);Anion exchanger 1(AE1)與Chloride channel 3(CLC-3)的染色位置。由NKA活性的測定實驗中可以發現,莫三比克吳郭魚馴養在去離子水環境下,鰓上NKA活性略高於馴養在原生的淡水環境下的吳郭魚,而馴養在海水環境下的吳郭魚鰓上NKA的活性則顯著高於淡水馴養組與去離子水馴養組的吳郭魚,由此可知馴養在去離子水與海水中的吳郭魚,較淡水組消耗更多的能量來推動二級主動運輸蛋白的運作,以維持體內滲透壓的恒定。由氯離子傳輸蛋白免疫染色發現,當吳郭魚馴養在去離子水與淡水的環境時,AE1與CLC-3均位於NKA免疫反應的細胞(NKA-immurnoreactive cell,簡稱NKIR細胞)基底膜上,NKCC位於NKIR細胞頂端的位置,CFTR則沒有反應。而當吳郭魚適應在海水環境時, NKCC 、AE1與CLC-3均位於NKIR細胞的基底膜上,而CFTR則出現在NKIR細胞的頂端。由此可以推論:當吳郭魚適應在淡水與去離子水環境時,NKCC將氯離子由外界帶入NKIR細胞中,AE1帶動血液中的氯離子進入NKIR細胞,CLC-3則將氯離子帶入血管中;當吳郭魚適應在海水中時,魚體內氯離子過多,則由CFTR將氯離子由NKIR細胞傳送至環境中,CLC-3氯離子由細胞帶至血管中,AE1與NKCC將氯離子由血管中傳送至細胞,藉由鰓上氯離子傳輸蛋白在不同環境下位於鰓表皮NKIR細胞內的不同極性分佈,來調控細胞內與體內的氯離子濃度恒定。

Mozambique tilapia (Oreochromis mossambicus) is a euryhaline teleost which can survive in fresh water and seawater. In this study, whole mount and immunofluorescent double staining followed by confocal laser scanning microscopic observation was performed to realize Cl- transport mechanisms in gills by immunolocalization of Cl- transporters, i.e., Na+/K+/2Cl- cotransporter (NKCC), cystic fibrosis transmembrane conductance regulator (CFTR), anion exchanger 1 (AE1), and chloride channel 3 (CLC-3) conterstained by Na+ / K+ -ATPase (NKA). The NKA activity in gills of DW-acclimated individuals was slightly higher than the FW-acclimated individuals, while the NKA activity in gills of SW-acclimated individuals was significantly higher than both FW- and DW- acclimated fish. The results indicated that DW- and SW-acclimated Mozambique tilapia required more energy to support the secondary active transport proteins for keeping homeostasis. The immunofluorescent double staining revealed that NKA as well as AE1 and CLC-3 were colocalized in the basolateral membrane, and NKCC were localized in the apical side of NKA immunoreactive cells(NKIR cells) of FW- or DW- acclimated individuals. NKA as well as AE1, CLC-3, and NKCC were colocalized in the basolateral membrane of NKIR cells of SW- acclimated individuals, while CFTR was localized only in the apical side of NKIR cells in SW-acclimated fish. In FW or DW group, NKCC transport Cl- into MR cells from environments ; AE1 transport Cl- into NKIR cells from the vessel, and CLC-3 transport Cl- into the vessel from NKIR cells. In SW- acclimated individuals, AE1 and NKCC ransport Cl- into NKIR cell from vessel, and CFTR transport extortionate Cl- to external environments. Taken together, differential expression of four Cl- transporters colocalized in the NKIR cells of gill epithelia implicated different mechanisms of Cl- transport among FW-, DW- or SW-acclimated tilapia to match the physiological requirements.
其他識別: U0005-1407200817302000
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